Novel Insights into the Synergistic Interaction of a Thioredoxin Reductase Inhibitor and TRAIL: The Activation of the ASK1-ERK-Sp1 Pathway
Figure 2
Treatment with a combination of DTCD and TRAIL activates apoptotic signal via the extrinsic and intrinsic pathways.
(A) Effects of DTCD plus TRAIL on mitochondrial membrane potential. A2780 cells were pretreated with(+)/without (-) z-IETD-fmk, and then were treated with DTCD (10 µM) and/or TRAIL (100 ng/mL). Mitochondrial membrane potential was measured by flow cytometry using DiOC6 dye. (B) Effects of TRAIL combined with DTCD on Caspase-8 and Bid cleavage. A2780 and SKOV3 cells were treated with DTCD (10 µM) and/or TRAIL (200 ng/mL) for the indicated time points, then subjected to Western blotting. (C) Levels of cytochrome c, Smac/DIABLO and Bax in cytosolic and pellet fractions were determined by Western blot. Following treatment, A2780 cells were lysed and cytosolic proteins were separated from mitochondria as described in Materials and Methods. COX IV and β-actin levels were used as internal controls for mitochondrial and cytosol specimen, respectively. (D) Effects of TRAIL combined with DTCD on levels of proapoptotic proteins. (E) DTCD combined with TRAIL regulates the expression of Bcl-2 and IAPs families.